CN111100804B - Lactobacillus paracasei LBP-YE01 wet powder, bacterial liquid and application thereof - Google Patents

Lactobacillus paracasei LBP-YE01 wet powder, bacterial liquid and application thereof Download PDF

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CN111100804B
CN111100804B CN201811266639.9A CN201811266639A CN111100804B CN 111100804 B CN111100804 B CN 111100804B CN 201811266639 A CN201811266639 A CN 201811266639A CN 111100804 B CN111100804 B CN 111100804B
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lactobacillus paracasei
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叶进荣
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Shenzhen Peptide Biotechnology Co ltd
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Abstract

The invention discloses wet powder of a lactobacillus paracasei strain LBP-YE01, a bacterial solution and application thereof, belonging to the field of microbial fermentation culture. The wet powder is prepared by the following method: inoculating Lactobacillus paracasei LBP-YE01 in thallus culture medium composed of semen glycines dregs, Oryza glutinosa and mineral water under 10-20 deg.C aseptic condition, culturing for 5 days, transferring into 0-5 deg.C aseptic environment, and culturing for 60 days to obtain mixture of bacteria and culture medium, i.e. wet powder. The bacterial liquid is prepared by the following method: transferring wet powder of lactobacillus paracasei LBP-YE01 into a sterile culture tank, adding mineral water and maltose, culturing at 42-45 deg.C for 48H, and filtering to obtain bacterial liquid. The wet powder and the bacterial liquid are used as nasal cavity cleaning liquid. The invention has the following advantages: can eliminate inflammation such as swelling of nasal mucosa, discharge pus and drain, ventilate and relieve pain; can be used for preventing viral common cold and asthma, and preventing recurrence.

Description

Lactobacillus paracasei LBP-YE01 wet powder, bacterial liquid and application thereof
Technical Field
The invention belongs to the field of microbial fermentation culture, and particularly relates to wet powder of lactobacillus paracasei strain LBP-YE01, a bacterial solution and application thereof.
Background
Allergic rhinitis refers to the reason that after an atopic individual is contacted with an allergen, IgE (immunoglobulin E) mediated mediator is mainly released, various immune active cells, cytokines and the like are involved in the non-infectious diseases of the nasal mucosa, and a non-specific antigen causes the organism to be immunized and atopic.
The vast majority of rhinitis virus infections are the leading cause, or bacterial and fungal infections are initiated on the basis of viral infections. More than 100 viruses are known to cause rhinitis, the most common being rhinovirus, influenza and parainfluenza, adenovirus, coronavirus, and myxovirus and paramyxovirus. The transmission mode is mainly through respiratory tract inhalation, and then pollutants or things enter the body. Other causes are nasal mucosal susceptibility, mainly the ability of antigenic substances to stimulate mast cells, basophils in mucosal tissues to cause release of chemical mediators.
Disclosure of Invention
The invention aims to disclose wet powder of lactobacillus paracasei strain LBP-YE 01.
The second purpose of the invention is to disclose the application of the wet powder of the lactobacillus paracasei strain LBP-YE 01.
The third purpose of the invention is to disclose a bacterial liquid of lactobacillus paracasei strain LBP-YE 01.
The fourth purpose of the invention is to disclose the application of the bacterial liquid of the lactobacillus paracasei strain LBP-YE 01.
The purpose of the invention is realized by the following technical scheme:
a wet powder of Lactobacillus paracasei strain LBP-YE01, wherein the wet powder of Lactobacillus paracasei strain LBP-YE01 is prepared by the following method: inoculating lactobacillus paracasei strain LBP-YE01 in a thallus culture medium consisting of soybean dregs, glutinous rice and mineral water under the aseptic condition of 10-20 ℃ for 5 days, and then transferring the mixture of the strain and the culture medium into the aseptic environment of 0-5 ℃ for 60 days, namely the wet powder of the lactobacillus paracasei strain LBP-YE 01;
the soybean dregs, the sticky rice and the mineral water are respectively in percentage by weight: 70% of soybean dregs, 20% of glutinous rice and 10% of mineral water.
The application of the wet powder of the lactobacillus paracasei strain LBP-YE01 in the technical scheme in the preparation of medicines for treating inflammation of nasal mucosa caused by virus bacteria, allergens and immune disorder.
The use of the above technical solution, wherein the wet powder of lactobacillus paracasei strain LBP-YE01 is used as a nasal cavity washing solution.
The bacterial liquid of the lactobacillus paracasei strain LBP-YE01, wherein the bacterial liquid of the lactobacillus paracasei strain LBP-YE01 is prepared by the following method: transferring wet powder of the lactobacillus paracasei strain LBP-YE01 of claim 1 into a sterile culture tank, adding mineral water and maltose, culturing at 42-45 ℃ for 48H, and filtering to obtain a bacterial solution of the lactobacillus paracasei strain LBP-YE 01;
the weight parts of the wet powder of the lactobacillus paracasei strain LBP-YE01, the mineral water and the maltose are 1 part of the wet powder of the lactobacillus paracasei strain LBP-YE01, 3-30 parts of the mineral water and 0.3-0.5 part of the maltose.
The bacterial liquid of the lactobacillus paracasei strain LBP-YE01 in the technical scheme is characterized in that the weight parts of the wet powder of the lactobacillus paracasei strain LBP-YE01, mineral water and maltose are 1 part of the wet powder of the lactobacillus paracasei strain LBP-YE01, 5 parts of mineral water and 0.5 part of maltose.
The bacterial liquid of the lactobacillus paracasei strain LBP-YE01 in the technical scheme, wherein the bacterial quantity in the bacterial liquid of the lactobacillus paracasei strain LBP-YE01 is one milliliter>1.5×108CFU/g。
The application of the lactobacillus paracasei strain LBP-YE01 bacterial liquid in the technical scheme in the preparation of medicines for treating inflammation of nasal mucosa caused by virus bacteria, allergens and immune disorder.
The application of the technical proposal, wherein the bacterial liquid of the lactobacillus paracasei strain LBP-YE01 is used as the nasal cavity cleaning liquid.
The preparation, isolation and identification of the lactobacillus paracasei strain LBP-YE01 of the present invention are described in the application documents of application No. 201810898195.4 entitled "lactobacillus paracasei strain LBP-YE01, its culture, its bacterial solution, a method for selecting and preserving a passaged strain, and its use" filed 8/8 in 2018; the preparation, isolation and characterization can also be carried out as follows:
1. sampling in the field, and selecting the trees or dead wood generating the flammulina velutipes in dark and humid places in the field forest of Heilongjiang province in China. About 100g of a mixture was collected from a mixed culture medium of the roots, bark and soil of Flammulina velutipes (the Lactobacillus paracasei strain of the present invention was not obtained every sampling, and thus a large amount was collected and selected.)
2. Preparing sample solution, placing the mixture in a 1L conical flask, adding 500ml mineral water, returning to the microorganism laboratory, stirring thoroughly (2H-3H) at room temperature about 25 deg.C, filtering with 200 mesh filter cloth to obtain filtrate in 1L conical flask, and placing 500ml filtrate in 100 deg.C water and shaking thoroughly for 15 min.
3. Culturing on an ultra-clean workbench, taking 1ml of filtrate, placing in an aseptic plate, pouring the prepared MRS agar culture medium with the temperature of about 48 ℃ into about 15ml of the plate, rotating the plate, mixing uniformly, and performing anaerobic culture for 72H in a constant-temperature incubator at 36 +/-1 ℃.
4. Selecting and purifying to observe colony morphology in a culture dish, and observing the morphology of lactobacillus paracasei in an MRS culture medium plate (as shown in figure 1), and after finding the colony, picking a small amount of thallus to a test tube MRS slant culture medium by using an inoculating loop.
5. The pure culture test tube MRS slant is inversely cultured for 72H in a constant temperature incubator at 36 +/-1 ℃ to obtain pure culture.
6. And (3) detecting and sending the obtained pure cultured test tube MRS slant to a special mechanism for strain determination, and evaluating whether the test tube MRS slant is a lactobacillus paracasei strain.
7. And (3) inoculating the obtained pure culture bacteria into a mixed culture medium containing 70% of soybean residue and 30% of glutinous rice flour by using an inoculating loop, culturing for 20 days at 10-20 ℃, and then culturing for about 180 days at 0-5 ℃.
Identification of Lactobacillus paracasei strain LBP-YE01 in accordance with the present invention:
1. morphological observation of the strains: a few bacteria on the MRS slant surface are picked by using an inoculating loop and placed on a glass slide, diluted by sterile water, covered by a cover glass and observed on a microscope with 3000-4000 times, and the result is shown in figure 2, wherein the bacteria are rod-shaped and have the length of about 10 mu m.
2. The applicant reserves the bacterial strain LBP-YE01 in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No. 15360; meanwhile, the G + C mol% content is measured by a microorganism research institute of Chinese academy of sciences, and a detection and identification report No. 224 of a micro index character (2018) issued by the microorganism research institute of Chinese academy of sciences indicates that: the G + C mol% of the genomic DNA of the test bacterial species (strain No.: LBP-YE01) was 45.28%. It can be seen that the G + C mol% of the strain of the invention is determined to be less than 3% -4% and at most not more than 5% of the same species of different strains with a similarity of 45.28%. Therefore, the report can indicate that the lactobacillus paracasei of the same species and different strains.
In order to ensure that each generation of strains has the same efficacy, the generation strains need to be selected and preserved, and the method for selecting and preserving the strain of the lactobacillus paracasei strain LBP-YE01 is as follows:
soaking 10mL of bacterial liquid from the lactobacillus paracasei strain LBP-YE01 bacterial liquid in a water bath at 100 ℃ for 5 minutes in an aseptic test tube, then transferring 1mL of bacterial liquid into an aseptic plate, pouring MRS agar culture medium cooled to 48 ℃ into about 15mL of the plate, rotating the plate, uniformly mixing, and then carrying out anaerobic culture for 72H in a constant temperature incubator at 36 +/-1 ℃;
after the culture is finished, taking out the plate, picking out a bacterial colony from a single bacterial colony by using an inoculating ring, inoculating 6-10 aseptic slant MRS agar culture medium test tubes by adopting a slant inoculation method, placing the slant of the inoculated test tube into a 36 +/-1 ℃ anaerobic culture medium for 72H, and preserving the strain by adopting a standard vacuum freeze drying preservation method;
the MRS agar culture medium is prepared by the following method: 10g of peptone, 5g of beef powder, 4g of yeast powder, 20g of glucose, 801 mL of tween and K2HPO4·7H2O2 g, sodium acetate 3H2O5 g, triammonium citrate 2g, MgSO4·7H2O 0.2g、MgSO4·4H2Adding 0.05g of O and 15g of agar powder into 1000mL of distilled water, heating to dissolve, adjusting the pH to 6.2 +/-0.2, subpackaging, and sterilizing at 121 ℃ for 15-20 min.
Slant inoculation method used in selection and preservation of passaged strains: slant inoculation is an inoculation method in which a small amount of strain is picked from a well-grown strain plate and transplanted onto a fresh slant culture medium. The slant inoculation method is often adopted in the strain enlargement culture and the strain preservation, and the inoculation tool used in the slant inoculation is an inoculating loop.
Vacuum freeze-drying preservation method used in selection and preservation of passage strain: the most effective method for preserving strains is to date, in which microorganisms are rapidly frozen at low temperature (-about 70 ℃) and then water is removed by sublimation under reduced pressure, so that the strains are always kept under low temperature, dry and anoxic conditions. The specific operation is as follows: the purity of the used strain needs to be noticed, the strain cannot be polluted by other bacteria, 2-3 ml of skimmed milk is sucked by a sterile suction pipe and added into a slant face of the strain to be preserved, the bacterial lawn is scraped by an inoculating loop and is gently stirred to be evenly suspended in the milk to form bacterial suspension, and the agar is not scraped into the milk.
The invention has the following beneficial effects:
1. the wet powder and the bacterial liquid have obvious curative effects on various inflammations of nasal mucosa caused by virus, bacteria, allergens and immune disorder, such as atrophic rhinitis, including chronic rhinitis, acute rhinitis, nasopharynx inflammation, burning sensation, frequent sneezing due to nasal itching, nosebleed and the like, have no pain, are quick in effect, and can quickly play roles in eliminating inflammations of nasal mucosa swelling and the like, discharging pus, draining, ventilating and relieving pain.
2. The wet powder and the bacterial liquid have exact curative effects on preventing virus cold and asthma, are convenient to use, safe and reliable, have no toxic or side effect, and are not easy to relapse after healing.
Description of the drawings:
1. FIG. 1 is a colony map of Lactobacillus paracasei in a MRS medium plate.
2. FIG. 2 shows the morphology of Lactobacillus paracasei observed on a microscope at 3000 to 4000 times.
Strain preservation information: the Lactobacillus paracasei strain LBP-YE01 is classified and named as Lactobacillus paracasei, and is already preserved in China general microbiological culture Collection center (CGMCC), wherein the preservation unit address is No. 3 of Xilu No.1 of Beijing Korean district, the preservation number is CGMCC No.15360, and the preservation date is 2018, 2 months and 27 days.
The specific implementation mode is as follows:
in order to facilitate the understanding of the technical scheme of the invention, the wet powder of lactobacillus paracasei strain LBP-YE01, the bacterial liquid and the application thereof are further described in the following by combining specific examples.
Example 1:preparation of wet powder of Lactobacillus paracasei strain LBP-YE 01:
weighing 70g of soybean dregs, 20g of glutinous rice and 10g of mineral water, and uniformly mixing to prepare a thallus culture medium; inoculating Lactobacillus paracasei LBP-YE01 in the bacteria culture medium under the aseptic condition of 15 ℃, transferring the mixture of bacteria and culture medium obtained by culturing for 5 days in the aseptic environment of 1 ℃ for 60 days, namely the wet powder of Lactobacillus paracasei LBP-YE 01.
Example 2:lactobacillus paracasei strainPreparation of LBP-YE01 culture:
this example is the same as example 1, except that: inoculating Lactobacillus paracasei LBP-YE01 in the bacteria culture medium under the aseptic condition of 10 ℃, transferring the mixture of bacteria and culture medium obtained by culturing for 5 days in the aseptic environment of 5 ℃ for 60 days, namely the wet powder of Lactobacillus paracasei LBP-YE 01.
Example 3:preparation of a culture of Lactobacillus paracasei strain LBP-YE 01:
this example is the same as example 1, except that: inoculating Lactobacillus paracasei LBP-YE01 in the thallus culture medium under the aseptic condition of 20 ℃, culturing for 5 days, and transferring to the aseptic environment of 0 ℃ to culture for 60 days to obtain the mixture of the bacteria and the culture medium, namely the wet powder of the Lactobacillus paracasei LBP-YE 01.
Example 4:preparation of a bacterial solution of Lactobacillus paracasei LBP-YE 01:
transferring wet powder of the lactobacillus paracasei strain LBP-YE01 prepared in example 1 into a sterile culture tank, adding mineral water and maltose, culturing at 42 ℃ for 48H, and filtering to obtain a bacterial solution of the lactobacillus paracasei strain LBP-YE 01;
the weight relationship among the wet powder of the lactobacillus paracasei strain LBP-YE01, the mineral water and the maltose is 1kg of wet powder of the lactobacillus paracasei strain LBP-YE01, 5kg of mineral water and 0.5kg of maltose.
Example 5:preparation of a bacterial solution of Lactobacillus paracasei LBP-YE 01:
transferring wet powder of the lactobacillus paracasei strain LBP-YE01 prepared in example 2 into a sterile culture tank, adding mineral water and maltose, culturing at 45 ℃ for 48H, and filtering to obtain a bacterial solution of the lactobacillus paracasei strain LBP-YE 01;
the weight relationship among the wet powder of the lactobacillus paracasei strain LBP-YE01, the mineral water and the maltose is 1kg of wet powder of the lactobacillus paracasei strain LBP-YE01, 15kg of mineral water and 0.4kg of maltose.
Example 6:preparation of a bacterial solution of Lactobacillus paracasei LBP-YE 01:
transferring wet powder of the lactobacillus paracasei strain LBP-YE01 prepared in example 3 into a sterile culture tank, adding mineral water and maltose, culturing at 44 ℃ for 48H, and filtering to obtain a bacterial solution of the lactobacillus paracasei strain LBP-YE 01;
the weight parts of the wet powder of the lactobacillus paracasei strain LBP-YE01, the mineral water and the maltose are 1kg of the wet powder of the lactobacillus paracasei strain LBP-YE01, 20kg of the mineral water and 0.3kg of the maltose.
The following specific experimental examples are provided to illustrate the beneficial effects of the bacterial solution of Lactobacillus paracasei LBP-YE01 on treating rhinitis:
the harmful bacteria in the bacteriostasis experiment of the invention are the most common harmful bacteria, and the nasal cavity can also inhale the harmful bacteria, such as staphylococcus aureus. The vast majority of the causes of rhinitis are caused by viral infection or bacterial or fungal infection of the mucous membrane in the nasal cavity after viral infection. The treatment principle of the wet powder of the lactobacillus paracasei strain LBP-YE01 and the bacterial liquid of the wet powder as the nasal cavity cleaning liquid is to kill and inhibit the infection of harmful bacteria (staphylococcus aureus, gram-negative bacteria, anaerobic coccus and the like) in the nasal cavity to the nasal cavity mucous membrane, protect beneficial bacteria to live in the nasal cavity, and further promote the immune system to normally work to achieve the effect of treating rhinitis.
Test example 1:and (3) testing the bacteriostatic performance of the lactobacillus paracasei LBP-YE01 bacterial liquid:
materials (I) and (II)
1. Strain:
escherichia coli CMCC44496, Salmonella CMCC50041, Staphylococcus aureus 26003, Listeria monocytogenes 54001, Bacillus cereus D1, Shigella sonnei ATCC29930, Enterobacter sakazakii CMCC45401, Pseudomonas aeruginosa CMCC10104
2. Culture medium:
the MRS culture medium is prepared by the following method: 10g of peptone, 5g of beef powder, 4g of yeast powder, 20g of glucose, 801 mL of tween and K2HPO4·7H2O2 g, sodium acetate 3H2O5 g, triammonium citrate 2g, MgSO4·7H2O0.2g、MgSO4·4H20.05g of O and 15g of agar powder are added into 1000mL of distilled waterHeating to dissolve, adjusting pH to 6.2 +/-0.2, subpackaging and sterilizing at 121 ℃ for 15-20 min.
Nutrient broth culture medium
The components: 10g of peptone, 3g of beef extract, 5g of sodium chloride and 1000mL of distilled water, and the pH value is 7.4.
3. Reagent:
pepsin/trypsin/papain/catalase (analytical grade), proteinase K (analytical grade), glucose (analytical grade), peptone/beef powder/yeast powder (biochemical reagent), dipotassium phosphate/triammonium citrate (analytical grade), sodium acetate (analytical grade), magnesium sulfate/tween 80 (analytical grade), manganese sulfate (analytical grade).
4. Main apparatus and equipment:
digital display acidimeter PH350 (Shanghai precision scientific instruments Co., Ltd.), biological safety cabinet BSC-1000A2 (Sujing group Antai Co., Ltd.), DNP-9272 type biochemical incubator (Shanghai Shenxian constant temperature plant), Agilent 1260 type high performance liquid chromatography (Agilent corporation of America), DSX-280B vertical pressure steam sterilizer (Shanghai Shenan medical instrument plant).
II, an experimental method:
1. pretreatment:
1.1, a device for indicating bacteria liquid:
the indicator bacteria are inoculated in a nutrient soup culture medium, cultured overnight at 37 ℃ and used for coating, and a bacteriostasis experiment is carried out.
1.2, preparation of test bacterium fermentation liquor:
lactobacillus paracasei LBP-YE01 was inoculated in MRS liquid medium and cultured at 37 ℃ for 24 hours to obtain fermentation broth for the experiment.
1.3, preparation of sterile fermentation filtrate of test bacteria:
centrifuging the fermentation liquid obtained in section 1.2 (4500r/min, 10min), filtering the supernatant with 0.22 μm microporous filter membrane for sterilization, and collecting the filtrate as sterile fermentation filtrate of test bacteria.
1.4, preparation of test bacteria:
and (3) washing the thallus precipitate obtained after the centrifugation in section 1.3 for 3 times by using sterile PBS, and then re-suspending to obtain the thallus suspension used for the experiment.
2. And (3) determining the antibacterial activity:
and (3) performing bacteriostatic activity determination on the fermentation liquor, the sterile fermentation filtrate and the thallus of the lactobacillus paracasei LEB-YE01 by using an oxford cup method, and evaluating the bacteriostatic effect according to the diameter of a bacteriostatic zone.
The indicator bacteria selected in the test are as follows: listeria monocytogenes 54001, Enterobacter sakazakii CMCC45401, Staphylococcus aureus 26003, Salmonella CMCC50041, Shigella sonnei ATCC29930, Bacillus cereus D1, Escherichia coli CMCC44496, and Pseudomonas aeruginosa CMCC 10104.
3. The antibacterial action of the sterile fermentation filtrate:
escherichia coli CMCC44496, Staphylococcus aureus 26003, Listeria monocytogenes 54001 and Shigella sonnei ATCC29930 are used as indicator bacteria, and are cultured by using sterile fermentation filtrate. In order to determine the influence of lactic acid produced during fermentation and the change of pH of MRS culture medium on the activity of pathogenic bacteria, the indicator bacteria were cultured with sterile fermentation filtrate (pH6.2), MRS liquid culture medium (supplemented with the same amount of lactic acid as in the sterile fermentation filtrate), and MRS liquid culture medium. The content of lactic acid in the sterile fermentation filtrate was determined by high performance liquid chromatography. After overnight culture, the indicator bacteria are inoculated into the four culture media in an inoculation amount with the volume fraction of 1%, and are cultured at constant temperature of 37 ℃, and are respectively sampled for 0 hour, 1 hour, 2 hours, 3 hours and 4 hours, and a tryptone soy agar culture medium is adopted for plate counting.
4. Influence of Heat treatment on the sterile fermentation filtrate or the fermentation broth:
the samples were treated in water baths at 60, 80 and 100 ℃ for 20min, high pressure steam at 121 ℃ for 20min, and a blank control was set. The method is characterized in that escherichia coli CMCC44496, shigella sonnei ATCC29930, staphylococcus aureus 26003 and Listeria monocytogenes 54001 are used as indicator bacteria, lactobacillus paracasei LBP-YE01 are used as test bacteria, and an oxford cup method is adopted for determining the bacteriostatic activity.
Thirdly, result and analysis:
1. and (3) determining the antibacterial activity:
as can be seen from Table 1, the fermented liquid and the sterile fermented filtrate of Lactobacillus paracasei LBP-YE01 have good inhibitory effects on all indicator bacteria including gram-positive bacteria and gram-negative bacteria, and the thallus has no inhibitory effect on all indicator bacteria. MRS liquid medium had no bacteriostatic effect (results not shown). The fermentation bacteria liquid has stronger bacteriostatic effect on all indicator bacteria than the sterile fermentation filtrate.
TABLE 1 Lactobacillus paracasei LBP-YE01 bacteriostatic activity assay
Figure BDA0001845031440000081
2. The antibacterial action of the sterile fermentation filtrate:
after lactobacillus paracasei LBP-YE01 is cultured in MRS liquid medium for 24h, the pH value of the MRS liquid medium is reduced from 6.2 to 3.62, and the concentration of lactic acid reaches 157.34 mM. In order to examine whether changes in the pH of lactic acid and MRS liquid medium produced during fermentation have an effect on the activity of pathogenic bacteria, the pathogenic bacteria were cultured with sterile fermentation filtrate (pH6.2) and MRS liquid medium (supplemented with 157.34mM lactic acid), respectively. After the fermentation filtrate is acted for 1h in the sterile fermentation filtrate, the viable count of Escherichia coli CMCC44496, Listeria monocytogenes 54001 and Shigella sonnei ATCC29930 is reduced by 1.7-1.9lgCFU/mL, and Staphylococcus aureus 26003 loses activity. When the pH value of the sterile fermentation filtrate is adjusted to 6.2, the viable count of pathogenic bacteria at the same time point has no significant difference (p is greater than 0.05) compared with that in MRS liquid, and the bacteriostatic activity of the sterile fermentation filtrate is related to the pH value of the sterile fermentation filtrate.
The viable count of pathogenic bacteria in MRS liquid culture medium supplemented with 157.34mM lactic acid is obviously reduced compared with that in MRS liquid culture medium, which indicates that lactic acid generated in the LBP fermentation process of lactobacillus paracasei is one of the factors for generating bacteriostatic activity.
MRS broth supplemented with 157.34mM lactic acid had a stronger inhibitory effect against four pathogens than the sterile fermentation filtrate, probably because the amount of non-dissociated lactic acid was greater in MRS broth than in the sterile fermentation filtrate.
3. Influence of heat treatment on bacteriostatic activity of the sterile fermentation filtrate:
the results in Table 2 show that the bacteriostatic activity of the sterile fermentation filtrate to pathogenic bacteria has no significant difference (P >0.05) compared with the control group after the sterile fermentation filtrate is treated under 4 temperature conditions, which indicates that the bacteriostatic substance generated by the Lactobacillus paracasei LBP-YE01 is not sensitive to heat.
TABLE 2 Effect of Heat treatment on bacteriostatic Activity of sterile fermentation filtrates
Figure BDA0001845031440000091
Fourthly, the experimental result shows that:
the fermentation liquor and the sterile fermentation filtrate of the lactobacillus paracasei LBP-YE01 have wide antibacterial spectrum and have better antibacterial activity on gram-positive bacteria and gram-negative bacteria. The generation of lactic acid and the reduction of the pH of the fermentation broth during the fermentation process are the main bacteriostatic factors. The antibacterial substance has thermal stability.
While the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims; meanwhile, any equivalent changes, modifications and variations of the above embodiments according to the essential technology of the present invention are within the scope of the technical solution of the present invention.

Claims (8)

1. A wet powder of Lactobacillus paracasei strain (Lactobacillus paracasei) LBP-YE01, which is prepared by the following method: inoculating lactobacillus paracasei strain LBP-YE01 in a thallus culture medium consisting of soybean dregs, glutinous rice and mineral water under the aseptic condition of 10-20 ℃ for 5 days, and then transferring the mixture of the strain and the culture medium into the aseptic environment of 0-5 ℃ for 60 days, namely the wet powder of the lactobacillus paracasei strain LBP-YE 01;
the soybean dregs, the sticky rice and the mineral water are respectively in percentage by weight: 70% of soybean dregs, 20% of glutinous rice and 10% of mineral water;
the lactobacillus paracasei strain LBP-YE01 is CGMCC No. 15360.
2. Use of wet powder of lactobacillus paracasei strain LBP-YE01 according to claim 1 for the preparation of a medicament for the treatment of inflammation of the nasal mucosa caused by viral bacterial, allergenic and immunological disorders.
3. Use according to claim 2, characterized in that: use of wet powder of Lactobacillus paracasei strain LBP-YE01 as a nasal wash.
4. The bacterial liquid of a lactobacillus paracasei strain LBP-YE01, which is characterized in that the bacterial liquid of the lactobacillus paracasei strain LBP-YE01 is prepared by the following method: transferring wet powder of the lactobacillus paracasei strain LBP-YE01 of claim 1 into a sterile culture tank, adding mineral water and maltose, culturing at 42-45 ℃ for 48H, and filtering to obtain a bacterial solution of the lactobacillus paracasei strain LBP-YE 01;
the weight parts of the wet powder of the lactobacillus paracasei strain LBP-YE01, the mineral water and the maltose are 1 part of the wet powder of the lactobacillus paracasei strain LBP-YE01, 3-30 parts of the mineral water and 0.3-0.5 part of the maltose.
5. The bacterial liquid of the lactobacillus paracasei strain LBP-YE01 according to claim 4, wherein: the weight parts of the wet powder of the lactobacillus paracasei strain LBP-YE01, the mineral water and the maltose are 1 part of the wet powder of the lactobacillus paracasei strain LBP-YE01, 5 parts of the mineral water and 0.5 part of the maltose.
6. The bacterial liquid of the Lactobacillus paracasei strain LBP-YE01 according to claim 4 or 5, wherein: the number of bacteria in the bacterial suspension of Lactobacillus paracasei LBP-YE01 per ml>1.5×108CFU/g。
7. Use of the bacterial liquid of the lactobacillus paracasei strain LBP-YE01 as claimed in any one of claims 4 to 6 for the preparation of a medicament for treating inflammation of nasal mucosa caused by viral bacteria, allergens and immunological disorders.
8. Use according to claim 7, characterized in that: the bacterial liquid of Lactobacillus paracasei LBP-YE01 is used as nasal cavity cleaning solution.
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Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104489687A (en) * 2015-01-08 2015-04-08 张鹏宴 Healthcare food containing lactobacillus paracasei for treating fatty liver

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CN104605345B (en) * 2015-01-08 2016-06-22 鲁东大学 The health food for treating hyperlipemia containing a kind of Lactobacillus paracasei

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104489687A (en) * 2015-01-08 2015-04-08 张鹏宴 Healthcare food containing lactobacillus paracasei for treating fatty liver

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